猫泛白细胞减少症病毒SH0918株的分离鉴定及遗传进化分析  

Isolation,Identifi cation and Genetic Evolution of Feline Panleucopenia Virus Strain SH0918

作　　者：林亮亮 吴赛 汤傲星 刘春草 谈晓梅 程松 梁留存 滕耀鹏 朱世强 朱杰[1] 李传锋 孟春春[1] 刘光清[1] LIN Liangliang;Wu Sai;TANG Aoxing;LIU Chuncao;TAN Xiaomei;CHENG Song;LIANG Liucun;TENG Yaopeng;ZHU Shiqiang;ZHU Jie;LI Chuanfeng;MENG Chunchun;LIU Guangqing(Team for Companion Animal Biosafety and Prevention Technology,Shanghai Institute of Veterinary Medicine,CAAS,Shanghai 200241,China;Nanjing Agricultural University,Nanjing 210095,China;Henan Institute of Science and Technology,Xinxiang 453003,China;Laboklin Laboratory for Clinical Diagnostics,Guangzhou 510700,China;Shenzhen Pingshan Hualing Animal Hospital,Shenzhen 518118,China)

机构地区：[1]中国农业科学院上海兽医研究所,上海200241 [2]南京农业大学,南京210095 [3]河南省科技学院,新乡453003 [4]纳博科临(广州)生物科技服务有限公司,广州510700 [5]深圳市坪山区花菱动物医院,深圳518118

出　　处：《中国动物传染病学报》2024年第5期152-161,共10页Chinese Journal of Animal Infectious Diseases

基　　金：上海市科技兴农项目(沪农科创字(2019)第3-3号);国家重点研发计划项目(2016YFD0500108、2016YFD0501003);上海市科技兴农创新项目(沪农科创字(2019)第3-3号);动物基因工程疫苗国家重点实验室开放课题(AGVSKL-ZD-202010)。

摘　　要：本文对2015—2022年收集的25份疑似猫瘟样本进行了PCR检测和病原分离,同时还根据测定的VP2基因序列,对本文分离的FPV流行毒株与国内外参考毒株的同源性以及遗传演化进行了分析。检测结果表明,在25份可疑样本中有6份FPV阳性样品。在对其VP2基因进行测序鉴定以后,我们重点对1株在F81细胞中增殖滴度较高的FPV(SH0918株)进行了分离和鉴定,并详细研究了该毒株的生物学特性。研究结果表明,SH0918株可以在F81中良好增殖,并产生特异性致病变效应,其TCID_(50)为10^(-5.11)/0.1mL;IFA的检测结果进一步证明在感染细胞中存在FPV的特异性蛋白表达;电镜检测结果表明,纯化的细胞培养物中存在典型的FPV病毒粒子;血凝试验结果表明,FPV能够凝集猪红细胞,血凝效价为1∶32。测定的FPV SH0918株的全基因组长5124 bp;序列比对分析结果显示,本文分离的毒株与参考毒株之间的核苷酸序列同源性为98.4%~100%;进化树结果表明:本研究中的FPV分离毒株基本属于同一分支,其中SH0918株与国内分离株JN-FPV-96、TZ-FPV-112的亲缘关系较近,但与欧洲分离株以及疫苗株亲缘关系较远。本研究内容一方面丰富了我国FPV的流行病学资料,另一方面对新型猫瘟疫苗的研发也具有一定的参考价值。In the present study,25 suspected cat plague samples collected from 2021 to 2022 were tested by PCR and subject to virus isolation.At the same time,the VP2 gene of the Feline parvovirus(FPV)isolates were sequenced and analyzed for their homology and genetic evolution with the domestic and foreign reference strains.The results showed that 6 out of 25 suspected samples were found FPV positive and their VP2 genes were sequenced.Subsequently,we focused on the isolation and characterization of the FPV SH0918 strain.The results showed that SH0918 strain proliferated well in F81 cells and produced cytopathogenic eff ect.The TCID_(50)was determined as 10^(5.11)/0.1mL and the presence of FPV protein was confi rmed in infected cells using immunofl uorescence assay.Typical FPV virus particles were observed under electron microscopy.The FPV SH0918 strain agglutinated pig red blood cells with the hemagglutination titer at 1:32.In addition,the whole genome of FPV SH0918 was sequenced with 5124 bp in length.The nucleotide sequence homology between the isolated strains and reference strains was 98.4%-100%.Phylogenetic tree analysis showed that the FPV strains in this study basically belonged to the same branch,among which the SH0918 strain was closely related to the domestic strains JN-FPV-96 and TZ-FPV-112 but far from the European strains and vaccine strains.Taken together,this study enriched the epidemiological data of FPV in China and provided a reference for development of a new feline plague vaccine.

关 键 词：猫细小病毒 分离鉴定 生物学特性 VP2基因 遗传进化分析 

分 类 号：S852.65[农业科学—基础兽医学]